No abstract available.
Paramyxoviridae Infections* ; Polymerase Chain Reaction* ; Reverse Transcription*

Laccase activity of Pleurotus ostreatus is significantly increased by the addition of apple pomace. Among various conditions, the best concentration of apple pomace and cultivation time for the production of laccase by P. ostreatus was 2.5% and 9 days, respectively. Reverse transcription polymerase chain reaction analyses of laccase isoenzyme genes, including pox1, pox3, pox4, poxc, poxa3, and poxa1b, revealed a clear effect of apple pomace on transcription induction. Our findings reveal that the use of apple pomace can be a model for the valuable addition of similar wastes and for the development of a solid-state fermenter and commercial production of oyster mushroom P. ostreatus.
Fungi* ; Laccase* ; Pleurotus* ; Polymerase Chain Reaction ; Reverse Transcription

Objective. To determine the diagnostic accuracy of self-collected snorted and spit saliva in detecting COVID-19 using RT-PCR (ssRT-PCR) and lateral flow antigen test (ssLFA) versus nasopharyngeal swab RT-PCR (npRT-PCR). Methods. One hundred ninety-seven symptomatic subjects for COVID-19 testing in a tertiary hospital underwent snort-spit saliva self-collection for RT-PCR and antigen testing and nasopharyngeal swab for RT-PCR as reference. Positivity rates, agreement, sensitivity, specificity, and likelihood ratios were estimated. Results. Estimated prevalence of COVID-19 using npRT-PCR was 9% (exact 95% CI of 5.5% - 14.1%). A higher positivity rate of 13% in the ssRT-PCR assay suggested possible higher viral RNA in the snort-spit samples. There was 92.9% agreement between ssRT-PCR and npRT-PCR (exact 95% CI of 88.4% to 96.1%; Cohen’s Kappa of 0.6435). If npRT-PCR will be assumed as reference standard, the estimated Sensitivity was 83.3% (exact 95% CI of 60.8% to 94.2%), Specificity 93.9% (exact 95% CI of 89.3% to 96.5%), Positive predictive value of 57.7% (exact 95% CI of 38.9% to 74.5%), Negative predictive value of 98.2% (exact 95% CI of 95% to 99.4%), positive likelihood ratio of 3.65 (95% CI of 7.37 to 24.9), negative likelihood ratio of 0.178 (95% CI of 0.063 to 0.499). There was 84.84% agreement (95% exact CI of 79.1% to 89.5%; Cohen’s Kappa of 0.2356) between ssLFAvs npRT-PCR, sensitivity of 38.9% (exact 95% CI of 20.3% to 61.4%), specificity of 89.4% (exact 95% CI of 84.1% to 93.1%), PPV of 26.9% (95% CI of 13.7% to 46.1%), NPV of 93.6% (exact 95% CI of 88.8% to 96.4%), LR+ of 3.67 (95% CI of 1.79 - 7.51), LR – of 0.68 (95% CI of 0.47 - 0.99). Conclusion. Our data showed that snort-spit saliva RT-PCR testing had acceptable diagnostic performance characteristics and can potentially be used as an alternative to the standard nasopharyngeal/oropharyngeal swab RT-PCR test for COVID-19 in certain situations. However, our data also showed that snort-spit saliva antigen testing using lateral flow assay did not offer acceptable performance.
Saliva ; SARS-CoV-2 ; Reverse Transcription ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUND: Kruppel-like factor 4 (KLF4) is a transcription factor that regulates a diverse array of cellular processes, including development, differentiation, proliferation, and apoptosis. Although its function in keratinocytes has been widely studied, its exact role in psoriasis has not been elucidated. OBJECTIVE: We designed this study to investigate epidermal expression levels of KLF4 and the change in KLF4 expression after treatment in patients with psoriasis. METHODS: We compared the expression levels of KLF4 in the basal, suprabasal, and superficial epidermal layers, in psoriatic lesional, non-lesional, and normal skin, using an immunoreactivity intensity distribution index (IRIDI). In addition, we measured the change in KLF4 expression on the basis of the IRIDI and by reverse transcription polymerase chain reaction (RT-PCR) analysis after treatment. RESULTS: The combined IRIDI scores in psoriatic lesional skin were significantly higher than the scores in both non-lesional and normal skin. The psoriatic epidermis, particularly the suprabasal layer, showed a significantly increased IRIDI score compared to that of non-lesional and normal skin, which was significantly decreased after treatment. RT-PCR analysis exhibited a slight increase in KLF4 mRNA expression level after treatment; however, this increase was not significant. CONCLUSION: These data indicate that KLF4 could regulate epidermal proliferation and differentiation. Moreover, we believe that KLF4 may play an important role in the physiological reaction to counteract abnormal differentiation and proliferation of keratinocytes.
Apoptosis ; Epidermis ; Humans ; Keratinocytes ; Polymerase Chain Reaction ; Psoriasis* ; Reverse Transcription ; RNA, Messenger ; Skin ; Transcription Factors

This study was purposed to investigate the expression of the growth-factor independence 1 (GFI1) in patients with leukemia and its clinical significance. Bone marrow mononuclear cells were obtained from 65 newly diagnosed leukemia patients including 24 acute myeloid leukemia (AML), 18 chronic myelogenous leukemia (CML), 6 acute lymphoblastic leukemia (ALL), 17 blast crisis of chronic myelogenous leukemia and 13 patients with iron deficiency anemia (IDA) were used as controls. The relative expression of gene gfi1 was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and taqman quantitative real-time reverse transcription polymerase chain reaction (QRT-PCR). The results showed that gene expression of gene gfi1 in leukemia patients was obviously higher than that in controls and the difference was statistically significant (p < 0.01), in which the expression of gene gfi1 in newly diagnosed CML patients was higher than that in newly diagnosed AML, newly diagnosed ALL, CML-BCP patients and the difference was significant (p < 0.01). Expression of gene gfi1 in lymphocytic blast crisis of CML was higher than that in nonlymphocytic blast crisis of CML, and the difference was significant. It is concluded that gene gfi1 may play an important role in leukemia, especially in CML incidence and progression. The high level expression of gene gfi1 may be participate in the development of lymphoma.
DNA-Binding Proteins ; genetics ; Gene Expression ; Humans ; Leukemia ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics ; Transcription, Genetic ; genetics

White coloration of the muscle of the giant river prawn (Macrobrachium rosenbergii) is a serious problem in China. The Macrobrachium rosenbergii Nodavirus (MrNV) has been confirmed to be the pathogen that causes this disorder. To develop a rapid, sensitive and specific technology for the detection of Macrobrachium rosenbergii Nodavirus isolated from China (MrNV-China), a reverse-transcription loop- mediated isothermal amplification assay combined with a lateral flow dipstick (RT-LAMP-LFD) assay method is described. A set of four primers and a labeled probe were designed specifically to recognize six distinct regions of the MrNV RNA2 gene. Results showed the sensitivity of the RT-LAMP-LFD assay was ten-times higher than the reverse-transcription loop-mediated isothermal amplification assay (RT-LAMP) with agarose gel electrophoresis. The assay was conducted with one-step amplification at 61°C in a single tube within 45 min. No product was generated from shrimps infected with other viruses, including DNA viruses (infectious hypodermal and hematopoietic necrosis virus (IHHNV); white spot syndrome virus (WSSV)) and RNA viruses (Taura syndrome virus (TSV); infectious myonecrosis virus (IMNV); yellow head virus (YHV)). Results were visualized by the LFD method. Therefore, the described rapid and sensitive assay is potentially useful for MrNV detection.
Animals ; Nodaviridae ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Palaemonidae ; virology ; Reverse Transcription

A retron is a bacterial retroelement that encodes an RNA gene and a reverse transcriptase (RT). The former, once transcribed, works as a template primer for reverse transcription by the latter. The resulting DNA is covalently linked to the upstream part of the RNA; this chimera is called multicopy single-stranded DNA (msDNA), which is extrachromosomal DNA found in many bacterial species. Based on the conserved features in the eight known msDNA sequences, we developed a detection method and applied it to scan National Center for Biotechnology Information (NCBI) RefSeq bacterial genome sequences. Among 16,844 bacterial sequences possessing a retron-type RT domain, we identified 48 unique types of msDNA. Currently, the biological role of msDNA is not well understood. Our work will be a useful tool in studying the distribution, evolution, and physiological role of msDNA.
Biotechnology ; Chimera ; DNA ; DNA, Single-Stranded* ; Genome, Bacterial* ; Retroelements ; Reverse Transcription ; RNA ; RNA-Directed DNA Polymerase

Here, we describe a uracil-DNA glycosylase (UNG)-treated reverse transcription loop-mediated isothermal amplification (uRT-LAMP) for the visual detection of all subtypes of avian influenza A virus (AIV). The uRT-LAMP assay can prevent unwanted amplification by carryover contamination of the previously amplified DNA, although the detection limit of the uRT-LAMP assay is 10-fold lower than that of the RT-LAMP without a UNG treatment. To the best of our knowledge, this is the first successful application of deoxyuridine triphosphate/UNG strategy in RT-LAMP for AIV detection, and the assay can be applied for the rapid, and reliable diagnosis of AIVs, even in contaminated samples.
Animals ; Deoxyuridine ; Diagnosis ; DNA ; Influenza in Birds* ; Limit of Detection ; Reverse Transcription* ; Uracil-DNA Glycosidase

PURPOSE: Since the discovery of estrogen receptor-beta(ER-beta, five C-terminal variants of ER-beta were identified. We designed this study to investigate the pattern and clinical implications of ER-betaand its splicing variants expression in normal and malignant mammary tissues. METHODS: Using reverse transcription polymerase chain reaction (RT-PCR), we examined the expression levels of ER-alpha and ER-betaand its five splicing variants (beta1, beta2, beta3, beta4, beta5) in 50 paired normal and cancer tissues. We measured the densities of RT-PCR products using Tina version 2.10 (Raytest, Germany). Firstly, the incidence and intensity of ER-alpha and ER-beta and its five splicing variants were compared. Then the expression of ER-betamRNA splicing variants was also analyzed with regard to the ER-alphaprotein expression measured by immuno-histochemical staining and the menopausal status of the patients. Chi-square test and paired samples t-test were used for statistical analysis. Differences were considered to be significant with a p-value of less than 0.05. RESULTS: The expression of ER-betamRNA variants in normal breast and cancer tissues were as follows: ER-beta2 (100%/100%), ER-beta4 (76%/74%), ER-beta5 (32%/58%), and ER-beta1 (14%/16%). ER-beta3 was not detected at all. In terms of intensity, we observed a significant decrease of ER-beta2 (P<0.001) and an increase of ER-beta5 (P=0.004) in the mRNA expression levels among breast cancers compared to the corresponding normal breast tissues. Compared to the corresponding normal tissues, a significant decrease of ER-beta2 in cancer tissues was observed in patients with ER-alpha-positive (P<0.001), with age over 50 (P=0.01), and under 50 (P=0.04) as well, but not in patients with ER-alpha-negative (P=0.48). ER-beta4 also significantly decreased in patients with ER-alpha-positive (P=0.004) and with age over 50 (P=0.07). ER-beta5 showed a significant increment only in patient aged over 50 (P=0.04). CONCLUSION: ER-alpha mRNA expression significantly increases but ER-beta mRNA expression decreases in the cancer tissues compared to the corresponding normal tissues. Among ER-beta variant forms, ER-beta2 is predominant in both normal and malignant mammary tissues and ER-beta4, ER-beta5, and ER-beta1 in descending order but ER-beta3 does not express in mammary tissues. The decrease of ER-beta2 and ER-beta4 expression is prominent in cancer tissue especially in ER-alpha-positive cancers, which suggests that ER-beta2 and ER-beta4 may possess a regulatory function in mammary carcinogenesis. Further investigations to verify the roles of ER-beta variants are mandatory.
Breast ; Breast Neoplasms ; Carcinogenesis ; Estrogens* ; Humans ; Incidence ; Polymerase Chain Reaction ; Receptors, Estrogen ; Reverse Transcription ; RNA, Messenger

BACKGROUND: The detection of total anti-hepatitis A virus (anti-HAV) immunoglobulin (Ig) and IgM is important for diagnosing acute hepatitis A. Our laboratory introduced new commercial automated chemiluminescence immunoassays (CLIAs) for use in addition to pre-existing automated CLIA. We evaluated the rate of agreement in the detection of total anti-HAV Ig and IgM in serum samples between two automated CLIAs. METHODS: We analyzed 181 samples those were submitted for testing at Kangbuk Samsung Medical Center. We analyzed the rate of agreement between the ADVIA Centaur XP (Siemens, Germany) and the MODULAR ANALYTICS E170 (Roche, Switzerland) analyzers. We performed reverse transcription (RT)-PCR when there was a discrepancy between the results from the two analyzers. RESULTS: The agreement rates between the ADVIA Centaur XP and the MODULAR ANALYTICS E170 for total anti-HAV Ig and IgM were 97.2% and 98.9%, respectively. Discrepant results were obtained in seven cases; all were found to be HAV-negative based on RT-PCR analysis. CONCLUSIONS: The total anti-HAV Ig and IgM results obtained using the two automated analyzers were comparable. However, in cases of equivocal results tested by the ADVIA Centaur XP for anti-HAV IgM, retesting and follow-up testing of samples are recommended.
Hepatitis A ; Hepatitis A Antibodies ; Hepatitis A virus ; Immunoassay ; Immunoglobulin M ; Immunoglobulins ; Luminescence ; Reverse Transcription ; Viruses